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Forum: 454 Pyrosequencing 09-05-2012, 02:11 AM
Replies: 13
Views: 3,319
Posted By mohdsos
Hi all, I will share my findings with...

Hi all,

I will share my findings with broken emulsion too, it's more to do with the tissue lyzer and they way you dispense you final volume into the wells, usually I used to dispense near the...
Forum: 454 Pyrosequencing 07-16-2012, 10:52 PM
Replies: 23
Views: 4,794
Posted By mohdsos
The maximum good enrichment I did was 33% and it...

The maximum good enrichment I did was 33% and it was good.
Forum: 454 Pyrosequencing 05-26-2012, 10:20 PM
Replies: 8
Views: 2,603
Posted By mohdsos
Guys would you dilute the 10^7 tube, or the...

Guys would you dilute the 10^7 tube, or the original tube, because I am trying to pool 6 libraries.
Forum: 454 Pyrosequencing 05-26-2012, 05:21 AM
Replies: 6
Views: 2,156
Posted By mohdsos
well I've targeted a 14% enrichment just to...

well I've targeted a 14% enrichment just to overcome the two oil cups and run the full ptp using one cup emulsion and everything was chilled. the library size or the fragmentation was from 900-2.5kb...
Forum: 454 Pyrosequencing 05-13-2012, 04:22 AM
Replies: 1
Views: 1,175
Posted By mohdsos
I did 33% and it went ok, the mixed beads was a...

I did 33% and it went ok, the mixed beads was a little bit high, but below 20% of the overall reads.

Good luck :rolleyes:
Forum: 454 Pyrosequencing 05-13-2012, 04:17 AM
Replies: 6
Views: 2,156
Posted By mohdsos
a Great 454 Run

Hi all,

I've made a great run, i will share my mods when then output is validated.



http://dl.dropbox.com/u/619938/readlength_GACT_summary%20%281%29.png...
Forum: Academic/Non-Profit Jobs 05-05-2012, 05:50 AM
Replies: 1
Views: 2,950
Posted By mohdsos
is the job available for international applicant.

is the job available for international applicant.
Forum: 454 Pyrosequencing 04-06-2012, 11:48 PM
Replies: 7
Views: 4,568
Posted By mohdsos
hi JKistler, you can quantify the amplicons...

hi JKistler,

you can quantify the amplicons and pool them with consideration of a normalized concentration of all, then you can purify then using ampure XP beads or gel cut.



mo
Forum: 454 Pyrosequencing 01-21-2012, 10:42 PM
Replies: 3
Views: 7,800
Posted By mohdsos
well, I am waiting for the upgrade, I am so doing...

well, I am waiting for the upgrade, I am so doing it. but the is a bump, ROCHE won't support that very run, as amplicons is not listed as FLX+ application, so in case that something went wrong, there...
Forum: 454 Pyrosequencing 07-25-2011, 05:54 AM
Replies: 3
Views: 1,240
Posted By mohdsos
yes I did. 13% , but I deposited 2M, yet in...

yes I did.
13% , but I deposited 2M, yet in the wells are ~700,000
I've just received the lib , I didn't prepare it.
Forum: 454 Pyrosequencing 07-25-2011, 02:53 AM
Replies: 3
Views: 1,240
Posted By mohdsos
Sediment in the substrate tube in the XLR70

Hi everyone,

I've had an issue with one previous run. I've received a normalized and amplified RNA library.
I've titrate the library and ended up with:

cpb enrichment
0.5 1.12%
2 ...
Forum: 454 Pyrosequencing 07-25-2011, 12:14 AM
Replies: 17
Views: 6,714
Posted By mohdsos
I would recommend double ampure beads selection....

I would recommend double ampure beads selection. or gel purification,

the dimer can cause your enrichment to be high. once I had more that 80% enrichment.


Mo
Forum: 454 Pyrosequencing 06-15-2011, 02:57 AM
Replies: 24
Views: 5,928
Posted By mohdsos
I had it once, I figured out that I had...

I had it once, I figured out that I had enrichment rate of 80% WOW. I think you need to check of primer dimer in case if it was amplicons. in general it must be copy number issue. do another emulsion...
Forum: 454 Pyrosequencing 12-19-2010, 10:12 PM
Replies: 6
Views: 4,114
Posted By mohdsos
I use a normal hood no air flow only light. and...

I use a normal hood no air flow only light. and it's fine .
Forum: 454 Pyrosequencing 12-19-2010, 04:32 AM
Replies: 0
Views: 1,497
Posted By mohdsos
Exclamation ptp pricing issue

Hi there I have a question it might be not technically related but I would prefer your help. I am dealing with a local procurement vendor and it seems I don't have a really representative price i...
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